Novel anti-inflammatory androstane derivative compositions

ABSTRACT

There is provided a crystalline chemical composition comprising a compound of formula (I)  
                 
 
     in which the crystal lattice is stabilised by the presence of a guest molecule, characterised in the crystalline composition is of space group P2 1 2 1 2 1  having unit cell dimensions of about 7.6±0.6 Å, 12.7±0.7 Å, and 33±3 Å when determined at 120K.

[0001] This application is a Continuation-in-part of U.S. patentapplication Ser. No. 10/200364 filed on Jul. 22, 2002 which is aContinuation-in-part of U.S. patent application Ser. No. 10/067010 filedon Feb. 4, 2002 which is a Continuation-in-part of U.S. patentapplication Ser. No. 09/958050 filed on Oct. 2, 2001, which is basedupon International Patent Application No. PCT.GB01.03495 filed Aug. 3,2001, which claims priority to United Kingdom Patent Application No. GB0019172.6 filed Aug. 5, 2000.

[0002] The present invention relates to a novel composition containingan anti-inflammatory and anti-allergic compound of the androstane seriesand to processes for its preparation. The present invention also relatesto pharmaceutical formulations containing the composition and totherapeutic uses thereof, particularly for the treatment of inflammatoryand allergic conditions.

[0003] Glucocorticoids which have anti-inflammatory properties are knownand are widely used for the treatment of inflammatory disorders ordiseases such as asthma and rhinitis. For example, U.S. Pat. No.4,335,121 discloses6α,9α-Difluoro-17α-(1-oxopropoxy)-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester (known by the generic name of fluticasonepropionate) and derivatives thereof. The use of glucocorticoidsgenerally, and especially in children, has been limited in some quartersby concerns over potential side effects. The side effects that arefeared with glucocorticoids include suppression of theHypothalamic-Pituitary-Adrenal (HPA) axis, effects on bone growth inchildren and on bone density in the elderly, ocular complications(cataract formation and glaucoma) and skin atrophy. Certainglucocorticoid compounds also have complex paths of metabolism whereinthe production of active metabolites may make the pharmacodynamics andpharmacokinetics of such compounds difficult to understand. Whilst themodern steroids are very much safer than those originally introduced, itremains an object of research to produce new molecules which haveexcellent anti-inflammatory properties, with predictable pharmacokineticand pharmacodynamic properties, with an attractive side effect profile,and with a convenient treatment regime.

[0004] We have now identified a novel glucocorticoid compound and acrystalline composition thereof which substantially meets theseobjectives.

[0005] Thus, according to one aspect of the invention, there is provideda crystalline chemical composition comprising a compound of formula (I)

[0006] in which the crystal lattice is stabilised by the presence of aguest molecule, characterised in the crystalline composition is of spacegroup P2₁2₁2₁ having unit cell dimensions of about 7.7±0.6 Å, 13.7±0.7Å, and 37±3 Å when determined at 120K (hereinafter “a composition of theinvention”).

[0007] In the composition of the invention the crystalline lattice isstabilised by a hydrogen bonding interaction between the hydrogen atomof the C11 hydroxy on the compound of formula (I) with the oxygen of theC3 carbonyl on a second molecule of the compound of formula (I).

[0008] The nature of the crystal lattice can be seen by reference toFIG. 1 which shows the spacial arrangement of 4 molecules of steroid and8 guests within a single unit cell for an example composition. Detail ofthe hydrogen bond interaction between the steroid molecules is shown inFIG. 3. FIG. 2 shows the conformation of the steroid molecule with 2guest molecules present.

[0009] We have determined the XRPD profiles for a number of compositionsaccording to the invention. These XRPD profiles are also apparentlycharacteristic of the crystalline composition according to theinvention. In particular they exhibit one or more of the following 3features when determined at ambient temperature (eg around 295K):

[0010] (a) A peak in the range of around 4.3-5.0.

[0011] (b) A peak in the range of around 6.6-6.9.

[0012] (c) A peak in the range of around 11.5-11.8.

[0013] Typically they exhibit 2 or more of the above 3 features,especially 3 of the above features.

[0014] The XRPD profiles of compositions of the invention whencrystallographically pure also preferably exhibit one or more of thefollowing 6 features when determined at ambient temperature (eg around295K):

[0015] (a) Absence of a peak at around 7.3 (eg around 7.1-7.5) which isassociated with the profile of unsolvated Form 1, 2 and 3 polymorphs;

[0016] (b) Absence of a peak at around 7.5 (eg around 7.2-7.7) which isassociated with the profile of another class of compositions of compoundof formula (I);

[0017] (c) Absence of a peak at around 12.5 (eg around 12.3-12.7) whichis associated with the profile of unsolvated Form 1, 2 and 3 polymorphs;

[0018] (d) Absence of a peak at around 8.8-9.6 which is associated withthe profile of another class of compositions of compound of formula (I);

[0019] (e) Absence of a peak at around 10.5-11.1 which is associatedwith the profile of another class of compositions of compound of formula(I);

[0020] (f) Absence of a peak at around 12.2-12.6 which is associatedwith the profile of another class of compositions of compound of formula(I).

[0021] Preferably one or more preferably both of features (a) and (b) atleast are exhibited. Preferably 3 or more preferably 4, especially 5,most especially all 6 of the above 6 features are exhibited.

[0022] The chemical name of the compound of formula (I) is6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester.

[0023] The compound of formula (I) and compositions thereof havepotentially beneficial anti-inflammatory or anti-allergic effects,particularly upon topical administration, demonstrated by, for example,its ability to bind to the glucocorticoid receptor and to illicit aresponse via that receptor, with long acting effect. Hence, the compoundof formula (I) and compositions thereof is useful in the treatment ofinflammatory and/or allergic disorders, especially in once-per-daytherapy.

[0024] Space group P2₁2₁2₁ is characterised by angles of 90° beingpresent in each of the 3 axes.

[0025] We have discovered that the compound of formula (I) can form acrystalline composition of characteristic space group, unit celldimensions and crystalline structure as evidenced by X-ray diffractionwith a number of guest molecules.

[0026] The guest molecule preferably has a relative molecular weight inthe range 16 to 150, more preferably 60 to 130, especially 70 to 120.Preferably the guest molecule is a liquid at ambient temperature andpressure (eg 295K, 1.013×10⁵Pa). However guest molecules which are aliquid under pressure may also be capable of acting as a guest molecule(especially under pressurised conditions). Substances which are solidsat ambient temperature and pressure are also included.

[0027] Examples of suitable guest molecules include solvents e.g.:

[0028] Aromatic compounds eg benzene and substituted derivatives,especially benzene substituted with one or more (eg 1 or 2) halogen orC₁₋₄alkyl groups, for example toluene, o-xylene, m-xylene,fluorobenzene, chlorobenzene, ethylbenzene.

[0029] Preferred guest molecules are pharmaceutically acceptablesubstances and, as described below, compositions of the inventioncontaining them may be used in therapy. However even if the guestmolecule is not pharmaceutically acceptable then such compositions maybe useful in the preparation of other compositions containing compoundof formula (I), for example, other compositions of the inventioncontaining guest molecules that are pharmaceutically acceptable orcompound of formula (I) in unsolvated form.

[0030] The stoichiometry of the composition will usually be such thatthe ratio of compound to formula (I) to guest molecule, in molar terms,is 1:5-0.25, preferably 1:3.0-0.3, more preferably 1:2.2-0.6, especially1: 2.2-0.8.

[0031] Unusually a composition of the invention has a crystal structurewhich is quite distinct from that of compound of formula (I) in theabsence of a guest molecule, eg. the compound of formula (I) asunsolvated polymorph Form 1 which has a space group of P2₁ (i.e. two ofthe axis angles are 90°) and cell dimensions of 7.6, 14.1, 11.8Å whendetermined at 150K. Thus if the guest molecule is removed below athreshold level (which will differ from guest to guest) for example byheating (optionally at reduced pressure eg under vacuum) then thecrystal structure of the composition starts to break down and convertsto that of the structure of an unsolvated compound of formula (I),typically unsolvated polymorph Form 2 or 3.

[0032] Preferably the unit cell dimentions are about 7.7±0.4 Å, 13.8±0.3Å, and 36.8±3 Å when determined at 120K. Usually the unit celldimensions are about 7.7±0.2 Å, 13.8±0.2 Å, and 36.8±2.7 Å whendetermined at 120K.

[0033] Table 1 shows the unit cell dimensions and peak positions for anumber of example compositions: TABLE 1 Guest molecule Unit celldimensions Peak positions Toluene 7.8 13.7 34.2 4.9 6.8 11.8 m-xylene7.8 13.8 35.9 4.8 6.8 11.5 o-xylene ND ND ND 5.0 6.9 11.7 fluorobenzene7.7 13.9 38.7 ND ND ND ethylbenzene 7.7 13.8 37.8 ND ND ND chlorobenzene7.8 13.7 39.3 4.3 6.6 —

[0034] Compound (I) undergoes highly efficient hepatic metabolism toyield the 17-β carboxylic acid (X) as the sole major metabolite in ratand human in vitro systems. This metabolite has been synthesised anddemonstrated to be >1000 fold less active than the parent compound in invitro functional glucocorticoid assays.

[0035] This efficient hepatic metabolism is reflected by in vivo data inthe rat, which have demonstrated plasma clearance at a rate approachinghepatic blood flow and an oral bioavailability of <1%, consistent withextensive first-pass metabolism.

[0036] In vitro metabolism studies in human hepatocytes havedemonstrated that compound (I) is metabolised in an identical manner tofluticasone propionate but that conversion of (I) to the inactive acidmetabolite occurs approximately 5-fold more rapidly than withfluticasone propionate. This very efficient hepatic inactivation wouldbe expected to minimise systemic exposure in man leading to an improvedsafety profile.

[0037] Inhaled steroids are also absorbed through the lung and thisroute of absorption makes a significant contribution to systemicexposure. Reduced lung absorption could therefore provide an improvedsafety profile. Studies with compound (I) have shown significantly lowerexposure to compound (I) than with fluticasone propionate after drypowder delivery to the lungs of anaesthetised pigs.

[0038] An improved safety profile is believed to allow the compound offormula (I) to demonstrate the desired anti-inflammatory effects whenadministered once-per day. Once-per-day dosing is considered to besignificantly more convenient to patients than the twice-per day dosingregime that is normally employed for fluticasone propionate.

[0039] Examples of disease states in which the compound of formula (I)and compositions thereof have utility include skin diseases such aseczema, psoriasis, allergic dermatitis, neurodermatitis, pruritis andhypersensitivity reactions; inflammatory conditions of the nose, throator lungs such as asthma (including allergen-induced asthmaticreactions), rhinitis (including hayfever), nasal polyps, chronicobstructive pulmonary disease, interstitial lung disease, and fibrosis;inflammatory bowel conditions such as ulcerative colitis and Crohn'sdisease; and auto-immune diseases such as rheumatoid arthritis.

[0040] The compound of formula (I) may also have use in the treatment ofconjunctiva and conjunctivitis.

[0041] The composition of the invention is expected to be most useful inthe treatment of inflammatory disorders of the respiratory tract e.g.asthma, COPD and rhinitis particularly asthma and rhinitis.

[0042] It will be appreciated by those skilled in the art that referenceherein to treatment extends to prophylaxis as well as the treatment ofestablished conditions.

[0043] As mentioned above, the composition of the invention is useful inhuman or veterinary medicine, in particular as an anti-inflammatory andanti-allergic agent.

[0044] There is thus provided as a further aspect of the invention thecomposition of the invention for use in human or veterinary medicine,particularly in the treatment of patients with inflammatory and/orallergic conditions, especially for treatment once-per-day.

[0045] According to another aspect of the invention, there is providedthe use of the composition of the invention for the manufacture of amedicament for the treatment of patients with inflammatory and/orallergic conditions, especially for treatment once-per-day.

[0046] In a further or alternative aspect, there is provided a methodfor the treatment of a human or animal subject with an inflammatoryand/or allergic condition, which method comprises administering to saidhuman or animal subject an effective amount of the composition of theinvention, especially for administration once-per-day.

[0047] The composition of the invention may be formulated foradministration in any convenient way, and the invention therefore alsoincludes within its scope pharmaceutical compositions comprising thecomposition of the invention together, if desirable, in admixture withone or more physiologically acceptable diluents or carriers.Pharmaceutical compositions suitable for once-per-day administration areof particular interest.

[0048] Further, there is provided a process for the preparation of suchpharmaceutical compositions which comprises mixing the ingredients.

[0049] The composition of the invention may, for example, be formulatedfor oral, buccal, sublingual, parenteral, local or rectaladministration, especially local administration.

[0050] Local administration as used herein, includes administration byinsufflation and inhalation. Examples of various types of preparationfor local administration include ointments, lotions, creams, gels,foams, preparations for delivery by transdermal patches, powders,sprays, aerosols, capsules or cartridges for use in an inhaler orinsufflator or drops (e.g. eye or nose drops), solutions/suspensions fornebulisation, suppositories, pessaries, retention enemas and chewable orsuckable tablets or pellets (e.g. for the treatment of aphthous ulcers)or liposome or microencapsulation preparations.

[0051] Advantageously compositions for topical administration to thelung include dry powder compositions and spray compositions.

[0052] Dry powder compositions for topical delivery to the lung byinhalation may, for example, be presented in capsules and cartridges foruse in an inhaler or insufflator of, for example, gelatine. Formulationsgenerally contain a powder mix for inhalation of the compound of theinvention and a suitable powder base (carrier substance) such as lactoseor starch. Use of lactose is preferred. Each capsule or cartridge maygenerally contain between 20 μg-10 mg of the compound of formula (I) ina composition of the invention optionally in combination with anothertherapeutically active ingredient. Alternatively, the composition of theinvention may be presented without excipients. Packaging of theformulation may be suitable for unit dose or multi-dose delivery. In thecase of multi-dose delivery, the formulation can be pre-metered (e.g. asin Diskus, see GB 2242134 or Diskhaler, see GB 2178965, 2129691 and2169265) or metered in use (e.g. as in Turbuhaler, see EP 69715). Anexample of a unit-dose device is Rotahaler (see GB 2064336). The Diskusinhalation device comprises an elongate strip formed from a base sheethaving a plurality of recesses spaced along its length and a lid sheethermetically but peelably sealed thereto to define a plurality ofcontainers, each container having therein an inhalable formulationcontaining a composition of the invention preferably combined withlactose. Preferably, the strip is sufficiently flexible to be wound intoa roll. The lid sheet and base sheet will preferably have leading endportions which are not sealed to one another and at least one of thesaid leading end portions is constructed to be attached to a windingmeans. Also, preferably the hermetic seal between the base and lidsheets extends over their whole width. The lid sheet may preferably bepeeled from the base sheet in a longitudinal direction from a first endof the said base sheet.

[0053] Pharmaceutical formulations which are non-pressurised and adaptedto be administered as a dry powder topically to the lung via the buccalcavity (especially those which are free of excipient or are formulatedwith a diluent or carrier such as lactose or starch, most especiallylactose) are of particular interest.

[0054] Spray compositions for topical delivery to the lung by inhalationmay for example be formulated as aqueous solutions or suspensions or asaerosols delivered from pressurised packs, such as a metered doseinhaler, with the use of a suitable liquefied propellant. Aerosolcompositions suitable for inhalation can be either a suspension or asolution and generally contain the composition of the inventionoptionally in combination with another therapeutically active ingredientand a suitable propellant such as a fluorocarbon or hydrogen-containingchlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes,especially 1,1,1,2-tetrafluoroethane,1,1,1,2,3,3,3-heptafluoro-n-propane or a mixture thereof. The aerosolcomposition may optionally contain additional formulation excipientswell known in the art such as surfactants e.g. oleic acid or lecithinand cosolvents e.g. ethanol. One example formulation is excipient freeand consists essentially of (e.g. consists of) composition of theinvention (optionally together with a further active ingredient) and apropellant selected from 1,1,1,2-tetrafluoroethane,1,1,1,2,3,3,3-heptafluoro-n-propane and mixture thereof. Another exampleformulation comprises particulate composition of the invention, apropellant selected from 1,1,1,2-tetrafluoroethane,1,1,1,2,3,3,3-heptafluoro-n-propane and mixture thereof and a suspendingagent which is soluble in the propellant e.g. an oligolactic acid orderivative thereof as described in WO94/21229. The preferred propellantis 1,1,1,2-tetrafluoroethane. Pressurised formulations will generally beretained in a canister (e.g. an aluminium canister) closed with a valve(e.g. a metering valve) and fitted into an actuator provided with amouthpiece.

[0055] Medicaments for administration by inhalation desirably have acontrolled particle size. The optimum particle size for inhalation intothe bronchial system is usually 1-10 μm, preferably 2-5 μm. Particleshaving a size above 20 μm are generally too large when inhaled to reachthe small airways. To achieve these particle sizes the particles of thecomposition of the invention as produced may be size reduced byconventional means e.g. by micronisation. The desired fraction may beseparated out by air classification or sieving. Preferably, theparticles will be crystalline, prepared for example by a process whichcomprises mixing in a continuous flow cell in the presence of ultrasonicradiation a flowing solution of compound of formula (I) as medicament ina liquid solvent with a flowing liquid antisolvent for said medicament(e.g. as described in International Patent Application PCT/GB99/04368)or else by a process which comprises admitting a stream of solution ofthe substance in a liquid solvent and a stream of liquid antisolvent forsaid substance tangentially into a cylindrical mixing chamber having anaxial outlet port such that said streams are thereby intimately mixedthrough formation of a vortex and precipitation of crystalline particlesof the substance is thereby caused (e.g. as described in InternationalPatent Application PCT/GB00/04327).

[0056] When an excipient such as lactose is employed, generally, theparticle size of the excipient will be much greater than the inhaledmedicament within the present invention. When the excipient is lactoseit will typically be present as milled lactose, wherein not more than85% of lactose particles will have a MMD of 60-90 μm and not less than15% will have a MMD of less than 15 μm.

[0057] Formulations for administration topically to the nose (e.g. forthe treatment of rhinitis) include pressurised aerosol formulations andaqueous formulations administered to the nose by pressurised pump.Formulations which are non-pressurised and adapted to be administeredtopically to the nasal cavity are of particular interest. Theformulation preferably contains water as the diluent or carrier for thispurpose. Aqueous formulations for administration to the lung or nose maybe provided with conventional excipients such as buffering agents,tonicity modifying agents and the like. Aqueous formulations may also beadministered to the nose by nebulisation.

[0058] Other possible presentations include the following:

[0059] Ointments, creams and gels, may, for example, be formulated withan aqueous or oily base with the addition of suitable thickening and/orgelling agent and/or solvents. Such bases may thus, for example, includewater and/or an oil such as liquid paraffin or a vegetable oil such asarachis oil or castor oil, or a solvent such as polyethylene glycol.Thickening agents and gelling agents which may be used according to thenature of the base include soft paraffin, aluminium stearate,cetostearyl alcohol, polyethylene glycols, woolfat, beeswax,carboxypolymethylene and cellulose derivatives, and/or glycerylmonostearate and/or non-ionic emulsifying agents.

[0060] Lotions may be formulated with an aqueous or oily base and willin general also contain one or more emulsifying agents, stabilisingagents, dispersing agents, suspending agents or thickening agents.

[0061] Powders for external application may be formed with the aid ofany suitable powder base, for example, talc, lactose or starch. Dropsmay be formulated with an aqueous or non-aqueous base also comprisingone or more dispersing agents, solubilising agents, suspending agents orpreservatives.

[0062] If appropriate, the formulations of the invention may be bufferedby the addition of suitable buffering agents.

[0063] The proportion of the active compound of formula (I) in the localcompositions according to the invention depends on the precise type offormulation to be prepared but will generally be within the range offrom 0.001 to 10% by weight. Generally, however for most types ofpreparations advantageously the proportion used will be within the rangeof from 0.005 to 1% and preferably 0.01 to 0.5%. However, in powders forinhalation or insufflation the proportion used will usually be withinthe range of from 0.1 to 5%.

[0064] Aerosol formulations are preferably arranged so that each metereddose or “puff” of aerosol contains 1 μg-2000 μg e.g. 20 μg-2000 μg,preferably about 20 μg-500 μg of compound of formula (I) optionally incombination with another therapeutically active ingredient.Administration may be once daily or several times daily, for example 2,3, 4 or 8 times, giving for example 1, 2 or 3 doses each time.Preferably the composition of the invention is delivered once or twicedaily. The overall daily dose with an aerosol will typically be withinthe range 10 μg-10 mg e.g. 100 μg-10 mg preferably, 200 μg-2000 μg.

[0065] Topical preparations may be administered by one or moreapplications per day to the affected area; over skin areas occlusivedressings may advantageously be used. Continuous or prolonged deliverymay be achieved by an adhesive reservoir system.

[0066] For internal administration the compound according to theinvention may, for example, be formulated in conventional manner fororal, parenteral or rectal administration. Formulations for oraladministration include syrups, elixirs, powders, granules, tablets andcapsules which typically contain conventional excipients such as bindingagents, fillers, lubricants, disintegrants, wetting agents, suspendingagents, emulsifying agents, preservatives, buffer salts, flavouring,colouring and/or sweetening agents as appropriate. Dosage unit formsare, however, preferred as described below.

[0067] Preferred forms of preparation for internal administration aredosage unit forms i.e. tablets and capsules. Such dosage unit formscontain from 0.1 mg to 20 mg preferably from 2.5 to 10 mg of thecompound of formula (I).

[0068] The compound according to the invention may in general may begiven by internal administration in cases where systemic adreno-corticaltherapy is indicated.

[0069] In general terms preparations, for internal administration maycontain from 0.05 to 10% of the active ingredient dependent upon thetype of preparation involved. The daily dose may vary from 0.1 mg to 60mg, e.g. 5-30 mg, dependent on the condition being treated, and theduration of treatment desired.

[0070] Slow release or enteric coated formulations may be advantageous,particularly for the treatment of inflammatory bowel disorders.

[0071] Since the compound of formula (I) is long-acting, preferably thecomposition of the invention will be delivered once-per-day and the dosewill be selected so that the compound has a therapeutic effect in thetreatment of respiratory disorders (e.g. asthma or COPD, particularlyasthma) over 24 hours or more.

[0072] The pharmaceutical compositions according to the invention mayalso be used in combination with another therapeutically active agent,for example, a β₂ adrenoreceptor agonist, an anti-histamine or ananti-allergic. The invention thus provides, in a further aspect, acombination comprising the composition of the invention together withanother therapeutically active agent, for example, a β₂-adrenoreceptoragonist, an anti-histamine or an anti-allergic.

[0073] Examples of β₂-adrenoreceptor agonists include salmeterol (e.g.as racemate or a single enantiomer such as the R-enantiomer),salbutamol, formoterol, salmefamol, fenoterol or terbutaline and saltsthereof, for example the xinafoate salt of salmeterol, the sulphate saltor free base of salbutamol or the fumarate salt of formoterol.Pharmaceutical compositions employing combinations with long-actingβ₂-adrenoreceptor agonists (e.g. salmeterol and salts thereof) areparticularly preferred, especially those which have a therapeutic effect(e.g. in the treatment of asthma or COPD, particularly asthma) over 24hours or more.

[0074] Since the compound of formula (I) is long-acting, preferably thecomposition comprising the compound of formula (I) and the long-actingβ₂-adrenoreceptor agonists will be delivered once-per-day and the doseof each will be selected so that the composition has a therapeuticeffect in the treatment of respiratory disorders effect (e.g. in thetreatment of asthma or COPD, particularly asthma) over 24 hours or more.

[0075] Examples of anti-histamines include methapyrilene or loratadine.

[0076] Other suitable combinations include, for example, otheranti-inflammatory agents e.g. NSAIDs (e.g. sodium cromoglycate,nedocromil sodium, PDE4 inhibitors, leukotriene antagonists, iNOSinhibitors, tryptase and elastase inhibitors, beta-2 integrinantagonists and adenosine 2a agonists)) or antiinfective agents (e.g.antibiotics, antivirals).

[0077] Also of particular interest is use of the composition of theinvention in combination with a phosphodiesterase 4 (PDE4) inhibitore.g. cilomilast or a salt thereof.

[0078] The combination referred to above may conveniently be presentedfor use in the form of a pharmaceutical formulation and thuspharmaceutical formulations comprising a combination as defined abovetogether with a physiologically acceptable diluent or carrier representa further aspect of the invention.

[0079] The compound according to the invention in combination withanother therapeutically active ingredient as described above may beformulated for administration in any convenient way, and the inventiontherefore also includes within its scope pharmaceutical formulationscomprising the composition of the invention in combination with anothertherapeutically active ingredient together, if desirable, in admixturewith one or more physiologically acceptable diluents or carriers. Thepreferred route of administration for inflammatory disorders of therespiratory tract will generally be administration by inhalation.

[0080] Further, there is provided a process for the preparation of suchpharmaceutical compositions which comprises mixing the ingredients.

[0081] Therapeutic agent combinations may be in any form, for examplecombinations may comprise a single dose containing separate particles ofindividual therapeutics, and optionally excipient material(s),alternatively, multiple therapeutics may be formed into individualmulticomponent particles, formed for example by coprecipitation, andoptionally containing excipient material(s).

[0082] The individual compounds of such combinations may be administeredeither sequentially in separate pharmaceutical compositions as well assimultaneously in combined pharmaceutical formulations. Appropriatedoses of known therapeutic agents will be readily appreciated by thoseskilled in the art.

[0083] The composition of the invention may be prepared by themethodology described hereinafter, constituting a further aspect of thisinvention.

[0084] A first process for preparing a composition of the inventioncomprises crystallising the composition from a solution containing acompound of formula (I) and the guest molecule. The solution containingthe guest molecule could be the guest itself when this a liquid, orcould be the guest dissolved in another liquid substance which substancedoes not act as a guest molecule.

[0085] Optionally, for better control and reproduceability, thecrystallisation process may be assisted by seeding with crystals of thecomposition of the invention. The seed crystals of the composition ofthe invention need not contain the same guest molecule.

[0086] A second process for preparing a composition of the inventioncomprises contacting the compound of formula (I) or a compositionaccording to the invention thereof in solid form with a liquidcontaining the guest molecule (for example by slurrying) and obtainingthe composition therefrom. The liquid containing the guest moleculecould be the guest itself when this a liquid, or could be the guestdissolved in another liquid substance which substance does not act as aguest molecule.

[0087] A third process for preparing a composition of the inventioncomprises contacting a compound of formula (I) or a compositionaccording to the invention thereof in solid form with a vapourcontaining the guest molecule. This process is suitable when the guesthas acceptable volatility e.g. when the guest is a solvent.

[0088] In the second and third processes, the compound of formula (I)may be employed in the form of a composition with a guest molecule or ina form without a guest molecule (eg as unsolvated polymorph Form 1, 2 or3). In the first process the compound of formula (I) or a compositionaccording to the invention may be dissolved in the solution or preparedin situ.

[0089] In one particular embodiment of this aspect of the invention theinput compound of formula (I) in the first, second and third processesis in the form of a substantially amorphous solid. Preferably thecompound of formula (I) in the form of a substantially amorphous solidis preferably in the form of substantially amorphous particles. Forexample the the compound of formula (I) in the form of substantiallyamorphous particles may be obtained by spray drying a solutioncontaining the compound of formula (I). Any solvent that will dissolvethe compound of formula (I) that can be evaporated safely in a spraydrying process may be used. Suitable solvents for forming the solutioninclude, but are not limited to, methyl acetate, ethyl acetate,isopropyl acetate, acetone, 2-butanone, 3-pentanone,4-methyl-2-pentanone, ethanol, methanol, 1-propanol, propan-2-ol,acetonitrile, chloroform, dichloromethane especially methylethylketone(2-butanone). Solution concentration will typically be 0.5-50%specifically 10-40% eg 20-30%. Lower concentrations may be more suitablefor preparing smaller particle sizes especially 2-4% e.g. 3.5-4%. Theconcentration that may be employed will be limited by the dissolutionpower of the solvent. Methylethylketone is preferred since it dissolvescompound of formula (I) at a relatively high concentration which resultsin production advantages. The compound of formula (I) may be employed innon-solvated form or in the form of a composition of the invention (e.g.with acetone). Preferably it is employed as the non-solvated Form 1polymorph. Spray drying maybe performed, for example, using apparatussupplied by Buchi or Niro. A pneumatic spray nozzle orifice of e.g. 0.04inches is suitable, although alternate atomization methods such asrotary and pressure nozzles can be used. Solution flow rate maytypically be in the range 1-100 ml/min, especially 15-30mi/min. Theinlet temperature and flow rate combination should be suitable toevaporate the solvent completely to minimize the risk of solvent trappedin the particle expediting an amorphous to crystalline transition. Inlettemperatures can range from 50-250° C., typically 100-200° C.

[0090] Compound of formula (I) in unsolvated form which is itself auseful substance has been found to exist in 3 crystalline polymorphicforms, Forms 1, 2 and 3, although Form 3 may be an unstable variant ofForm 2. The Forms are characterised by their XRPD patterns shown in FIG.8. Broadly speaking the Forms are characterised in their XRPD profilesby the absence of guest molecules and by peaks as follows:

[0091] Form 1: Peak at around 18.9 degrees 2Theta

[0092] Form 2: Peaks at around 18.4 and 21.5 degrees 2Theta

[0093] Form 3: Peaks at around 18.6 and 19.2 degrees 2Theta.

[0094] Forms 1 appears likely to be the thermodynamically most stableform since Forms 2 and 3 are converted into Form 1 on heating.

[0095] A process for preparing a compound of formula (I) as crystallineunsolvated Form 1 polymorph comprises dissolving compound of formula (I)in methylisobutylketone or ethyl acetate and producing compound offormula (I) as unsolvated Form 1 by addition of an anti-solvent such asiso-octane or toluene.

[0096] According to a first preferred embodiment of this process thecompound of formula (I) may be dissolved in ethyl acetate and compoundof formula (I) as unsolvated Form 1 polymorph may be obtained byaddition of toluene as anti-solvent. In order to improve the yield,preferably the ethyl acetate solution is hot and once the toluene hasbeen added the mixture is distilled to reduce the content of ethylacetate.

[0097] According to a second preferred embodiment of this process thecompound of formula (I) may be dissolved in methylisobutylketone andcompound of formula (I) as crystalline unsolvated Form 1 polymorph maybe obtained by addition of isooctane as anti-solvent.

[0098] A process for preparing a compound of formula (I) as unsolvatedForm 2 polymorph comprises dissolving compound of formula (I) inunsolvated form in methanol or dry dichloromethane and recrystallisingthe compound of formula (I) as unsolvated Form 2 polymorph. Typicallythe compound of formula (I) will be dissolved in hot methanol or drydichloromethane and allowed to cool.

[0099] A process for preparing a compound of formula (I) as unsolvatedForm 3 polymorph comprises dissolving compound of formula (I) inparticular as the composition with acetone in dichloromethane in thepresence of water (typically 1-3% water by volume) and recrystallisingthe compound of formula (I) as unsolvated Form 3 polymorph.

[0100] As mentioned above, compositions of the invention may also finduse as manufacturing intermediates in the preparation of compound offormula (I) in unsolvated form, or in the preparation of othercompositions of the invention, or in pharmaceutical compositionsthereof.

[0101] For example, a process for preparation of compound of formula (I)in unsolvated form (typically unsolvated polymorph Form 1) comprisesremoving the guest molecule from a composition of the invention.

[0102] A process for preparing a compound of formula (I) comprisesalkylation of a thioacid of formula (II)

[0103] or a salt thereof.

[0104] In this process the compound of formula (II) may be reacted witha compound of formula FCH₂L wherein L represents a leaving group (e.g. ahalogen atom, a mesyl or tosyl group or the like) for example, anappropriate fluoromethyl halide under standard conditions. Preferably,the fluoromethyl halide reagent is bromofluoromethane. Preferably thecompound of formula (II) is employed as a salt, particularly the saltwith diisopropylethylamine.

[0105] In a preferred process for preparing the compound of formula (I),the compound of formula (II) or a salt thereof is treated withbromofluoromethane optionally in the presence of a phase transfercatalyst. A preferred solvent is methylacetate, or more preferablyethylacetate, optionally in the presence of water. The presence of waterimproves solubility of both starting material and product and the use ofa phase transfer catalyst results in an increased rate of reaction.Examples of phase transfer catalysts that may be employed include (butare not restricted to) tetrabutylammonium bromide, tetrabutylammoniumchloride, benzyltributylammonium bromide, benzyltributylammoniumchloride, benzyltriethylammonium bromide, methyltributylammoniumchloride and methyltrioctylammonium chloride. THF has also successfullybeen employed as solvent for the reaction wherein the presence of aphase transfer catalyst again provides a significantly faster reactionrate. Preferably the product present in an organic phase is washedfirstly with aqueous acid e.g. dilute HCl in order to remove aminecompounds such as triethylamine and diisopropylethylamine and then withaqueous base e.g. sodium bicarbonate in order to remove any unreactedprecursor compound of formula (II).

[0106] Compounds of formula (II) may be prepared from the corresponding17(X-hydroxyl derivative of formula (III):

[0107] using for example, the methodology described by G. H. Phillippset al., (1994) Journal of Medicinal Chemistry, 37, 3717-3729. Forexample the step typically comprises the addition of a reagent suitablefor performing the esterification e.g. an activated derivative of2-furoic acid such as an activated ester or preferably a 2-furoyl halidee.g. 2-furoyl chloride (employed in at least 2 times molar quantityrelative to the compound of formula (III)) in the presence of an organicbase e.g. triethylamine. The second mole of 2-furoyl chloride reactswith the thioacid moiety in the compound of formula (III) and needs tobe removed e.g. by reaction with an amine such as diethylamine.

[0108] This method suffers disadvantages, however, in that the resultantcompound of formula (II) is not readily purified of contamination withthe by-product 2-furoyidiethylamide. We have therefore invented severalimproved processes for performing this conversion.

[0109] In a first such improved process we have discovered that by usinga more polar amine such as diethanolamine, a more water solubleby-product is obtained (in this case 2-furoyldiethanolamide) whichpermits compound of formula (II) or a salt thereof to be produced inhigh purity since the by-product can efficiently be removed by waterwashing.

[0110] Thus we provide a process for preparing a compound of formula(II) which comprises:

[0111] (a) reacting a compound of formula (III) with an activatedderivative of 2-furoic acid as in an amount of at least 2 moles of theactivated derivative per mole of compound of formula (III) to yield acompound of formula (IIA)

[0112] ; and

[0113] (b) removal of the sulphur-linked 2-furoyl moiety from compoundof formula (IIA) by reaction of the product of step (a) with an organicprimary or secondary amine base capable of forming a water soluble2-furoyl amide.

[0114] In two particularly convenient embodiments of this process wealso provide methods for the efficient purification of the end productwhich comprise either

[0115] (c1)when the product of step (b) is dissolved in a substantiallywater immiscible organic solvent, purifying the compound of formula (II)by washing out the amide by-product from step (b) with an aqueous wash,or

[0116] (c2) when the product of step (b) is dissolved in a watermiscible solvent, purifying the compound of formula (II) by treating theproduct of step (b) with an aqueous medium so as to precipitate out purecompound of formula (II) or a salt thereof.

[0117] In step (a) preferably the activated derivative of 2-furoic acidmay be an activated ester of 2-furoic acid, but is more preferably a2-furoyl halide, especially 2-furoyl chloride. A suitable solvent forthis reaction is ethylacetate or methylacetate (preferablymethylacetate) (when step (c1) may be followed) or acetone (when step(c2) may be followed). Normally an organic base e.g. triethylamine willbe present. In step (b) preferably the organic base is diethanolamine.The base may suitably be dissolved in a solvent e.g. methanol. Generallysteps (a) and (b) will be performed at reduced temperature e.g. between0 and 5° C. In step (c1) the aqueous wash may be water, however the useof brine results in higher yields and is therefore preferred. In step(c2) the aqueous medium is for example a dilute aqueous acid such asdilute HCl.

[0118] We also provide an alternative process for preparing a compoundof formula (II) which comprises:

[0119] (a) reacting a compound of formula (III) with an activatedderivative of 2-furoic acid in an amount of at least 2 moles ofactivated derivative per mole of compound of formula (III) to yield acompound of formula (IIA); and

[0120] (b) removal of the sulphur-linked 2-furoyl moiety from compoundof formula (IIA) by reaction of the product of step (a) with a furthermole of compound of formula (III) to give two moles of compound offormula (II).

[0121] In step (a) preferably the activated derivative of 2-furoic acidmay be an activated ester of 2-furoic acid, but is more preferably a2-furoyl halide, especially 2-furoyl chloride. A suitable solvent forhis step is acetone. Normally an organic base e.g. triethylamine will bepresent. In step (b) a suitable solvent is DMF or dimethylacetamide.Normally an organic base e.g. triethylamine will be present. Generallysteps (a) and (b) will be performed at reduced temperature e.g. between0 and 5° C. The product may be isolated by treatment with acid andwashing with water.

[0122] This aforementioned process is very efficient in that it does notproduce any furoylamide by-product (thus affording inter aliaenvironmental advantages) since the excess mole of furoyl moiety istaken up by reaction with a further mole of compound of formula (II) toform an additional mole of compound of formula (II).

[0123] Further general conditions for the conversion of compound offormula (III) to compound of formula (II) in the two processes justdescribed will be well known to persons skilled in the art.

[0124] According to a preferred set of conditions, however, we havefound that the compound of formula (II) may advantageously be isolatedin the form of a solid crystalline salt. The preferred salt is a saltformed with a base such as triethylamine, 2,4,6-trimethylpyridine,diisopropylethylamine or N-ethylpiperidine. Such salt forms of compoundof formula (II) are more stable, more readily filtered and dried and canbe isolated in higher purity than the free thioacid. The most preferredsalt is the salt formed with diisopropylethylamine. The triethylaminesalt is also of interest.

[0125] Compounds of formula (III) may be prepared in accordance withprocedures described in GB 2088877B.

[0126] Compounds of formula (III) may also be prepared by a processcomprising the following steps:

[0127] Step (a) comprises oxidation of a solution containing thecompound of formula (V). Preferably, step (a) will be performed in thepresence of a solvent comprising methanol, water, tetrahydrofuran,dioxan or diethylene glygol dimethylether. So as to enhance yield andthroughput, preferred solvents are methanol, water or tetrahydrofuran,and more preferably are water or tetrahydrofuran, especially water andtetrahydrofuran as solvent. Dioxan and diethylene glygol dimethyletherare also preferred solvents which may optionally (and preferably) beemployed together with water. Preferably, the solvent will be present inan amount of between 3 and 10vol relative to the amount of the startingmaterial (1 wt.), more preferably between 4 and 6 vol., especially 5vol. Preferably the oxidising agent is present in an amount of 1-9 molarequivalents relative to the amount of the starting material. Forexample, when a 50% w/w aqueous solution of periodic acid is employed,the oxidising agent may be present in an amount of between 1.1 and 10wt. relative to the amount of the starting material (1 wt.), morepreferably between 1.1 and 3 wt., especially 1.3 wt. Preferably, theoxidation step will comprise the use of a chemical oxidising agent. Morepreferably, the oxidising agent will be periodic acid or iodic acid or asalt thereof. Most preferably, the oxidising agent will be periodic acidor sodium periodate, especially periodic acid. Alternatively (or inaddition), it will also be appreciated that the oxidation step maycomprise any suitable oxidation reaction, e.g. one which utilises airand/or oxygen. When the oxidation reaction utilises air and/or oxygen,the solvent used in said reaction will preferably be methanol.Preferably, step (a) will involve incubating the reagents at roomtemperature or a little warmer, say around 25° C. e.g. for 2 hours. Thecompound of formula (IV) may be isolated by recrystallisation from thereaction mixture by addition of an anti-solvent. A suitable anti-solventfor compound of formula (IV) is water. Surprisingly we have discoveredthat it is highly desirable to control the conditions under which thecompound of formula (IV) is precipitated by addition of anti-solvente.g. water. When the recrystallisation is performed using chilled water(e.g. water/ice mixture at a temperature of 0-5° C.) although betteranti-solvent properties may be expected we have found that thecrystalline product produced is very voluminous, resembles a soft geland is very difficult to filter. Without being limited by theory webelieve that this low density product contains a large amount ofsolvated solvent within the crystal lattice. By contrast when conditionsof around 10° C. or higher are used (e.g. around ambient temperature) agranular product of a sand like consistency which is very easilyfiltered is produced. Under these conditions, crystallisation typicallycommences after around 1 hour and is typically completed within a fewhours (e.g. 2 hours). Without being limited by theory we believe thatthis granular product contains little or no solvated solvent within thecrystal lattice.

[0128] Step (b) will typically comprise the addition of a reagentsuitable for converting a carboxylic acid to a carbothioic acid e.g.using hydrogen sulphide gas together with a suitable coupling agent e.g.carbonyidiimidazole (CDl) in the presence of a suitable solvent e.g.dimethylformamide.

[0129] The advantages of the composition comprising a compound offormula (I) together with a guest compound according to the inventionmay include the fact that the substance appears to demonstrate excellentanti-inflammatory properties, with predictable pharmacokinetic andpharmacodynamic behaviour, with an attractive side-effect profile, longduration of action, and is compatible with a convenient regime oftreatment in human patients, in particular being amenable to once-perday dosing. Further advantages may include the fact that the substancehas desirable physical and chemical properties which allow for readymanufacture and storage. Alternatively it may serve as a usefulintermediate in the preparation of other forms of the compound offormula (I) or compositions thereof.

BRIEF DESCRIPTION OF THE FIGURES

[0130]FIG. 1: Figure showing the spacial arrangement of 4 steroid and 8guest molecules in the unit cell of a composition of the invention withtoluene (guest molecule darkened).

[0131]FIG. 2: Figure showing hydrogen bond interactions between steroidand guest for the composition of the invention with toluene

[0132]FIG. 3: Figure showing the spacial arrangement of 4 steroid and 8guest molecules in the unit cell of a composition of the invention withtoluene, and hydrogen bond interactions between two steroid molecules

[0133]FIG. 4: Enlarged XRPD profile of composition of the invention withtoluene

[0134]FIG. 5: Enlarged XRPD profile of composition of the invention withm-xylene

[0135]FIG. 6: Enlarged XRPD profile of composition of the invention witho-xylene

[0136]FIG. 7: Enlarged XRPD profile of composition of the invention withchlorobenzene

[0137]FIG. 8: XRPD profiles of unsolvated polymorphs 1, 2 and 3

[0138] The following non-limiting Examples illustrate the invention:

EXAMPLES General

[0139]¹H-nmr spectra were recorded at 400 MHz and the chemical shiftsare expressed in ppm relative to tetramethylsilane. The followingabbreviations are used to describe the multiplicities of the signals: s(singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd(doublet of doublets), ddd (doublet of doublet of doublets), dt (doubletof triplets) and b (broad). Biotage refers to prepacked silica gelcartridges containing KP-Sil run on flash 12i chromatography module.LCMS was conducted on a Supelcosil LCABZ+PLUS column (3.3 cm×4.6 mm ID)eluting with 0.1% HCO₂H and 0.01 M ammonium acetate in water (solventA), and 0.05% HCO₂H 5% water in acetonitrile (solvent B), using thefollowing elution gradient 0-0.7 min 0% B, 0.7-4.2 min 100% B, 4.2-5.3min 0% B, 5.3-5.5 min 0% B at a flow rate of 3 ml/min. The mass spectrawere recorded on a Fisons VG Platform spectrometer using electrospraypositive and negative mode (ES+ve and ES−ve).

[0140] The XRPD analyses shown in the figures were performed on

[0141] a) a Phillips X'pert MPD powder diffractometer, serial numberDY667. The pattern was recorded using the following acquisitionconditions: Tube anode: Cu, Start angle: 2.0°2θ, End angle: 45.0°2θ,Step size: 0.02°2θ, Time per step: 1 second. XRPD profiles werecollected at ambient temperature (295K) (FIG. 8);

[0142] b) a Philips PW1710 powder diffractometer. The pattern wasrecorded using the following acquisition conditions: Tube anode: Cu,Start angle: 3.5°2θ, End angle: 35.0°2θ, Step size: 0.02°2θ, Time perstep: 2.3 seconds. XRPD profiles were collected at ambient temperature(295K) (FIGS. 4-7);

[0143] The diffractometer used in each case can be determined by the endangle in the figure.

[0144] X-ray diffraction pattern collections referred to in Table 1 wereperformed in the following manners:

[0145] The crystal and molecular structures and corresponding unit celldimensions were determined from three-dimensional X-ray diffraction datacollected at 120+/−2 K. All measurements were made using a Bruker SMARTCCD diffractometer with graphite monochromated Mo-Kα radiation(λ=0.71073 Å) from a fine focus sealed tube source. The structure wassolved by direct methods and refined using full-matrix least-squaresprocedures which minimized the function Sw(Fo²-Fc²)². The Bruker SHELXsoftware was used throughout.

Intermediates Intermediate 1:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid diisopropylethylamine salt

[0146] A stirred suspension of6α,9α-difluoro-11β,17α-dihydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid (prepared in accordance with the procedure described in GB2088877B) (49.5 g) in methylacetate (500 ml) is treated withtriethylamine (35 ml) maintaining a reaction temperature in the range0-5° C. 2-Furoyl chloride (25 ml) is added and the mixture stirred at0-5° C. for 1 hour. A solution of diethanolamine (52.8 g) in methanol(50 ml) is added and the mixture stirred at 0-5° C. for at least 2hours. Dilute hydrochloric acid (approx 1 M, 550 ml) is addedmaintaining a reaction temperature below 15° C. and the mixture stirredat 15° C. The organic phase is separated and the aqueous phase is backextracted with methyl acetate (2×250 ml). All of the organic phases arecombined, washed sequentially with brine (5 ×250 ml) and treated withdi-isopropylethylamine (30 ml). The reaction mixture is concentrated bydistillation at atmospheric pressure to an approximate volume of 250 mland cooled to 25-30° C. (crystallisation of the desired product normallyoccurs during distillation/subsequent cooling). Tertiary butyl methylether (TBME) (500 ml) is added, the slurry further cooled and aged at0-5° C. for at least 10 minutes. The product is filtered off, washedwith chilled TBME (2×200 ml) and dried under vacuum at approximately40-50° C. (75.3 g, 98.7%). NMR (CDCl₃) δ: 7.54-7.46 (1H, m), 7.20-7.12(1H, dd), 7.07-6.99 (1H, dd), 6.48-6.41 (2H, m), 6.41-6.32 (1H, dd),5.51-5.28 (1H, dddd ²J_(H-F) 50 Hz), 4.45-4.33(1H, bd), 3.92-3.73 (3H,bm), 3.27-3.14 (2H, q), 2.64-2.12 (5H, m), 1.88-1.71 (2H, m), 1.58-1.15(3H, s), 1.50-1.38 (15H, m), 1.32-1.23 (1H, m), 1.23-1.15 (3H s),1.09-0.99 (3H, d)

Intermediate 2:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0147] Unsolvated Form 1

[0148] A mobile suspension of Intermediate 1 (12.61 g, 19.8 mmol) inethyl acetate (230 ml) and water (50 ml) is treated with a phasetransfer catalyst (benzyltributylammonium chloride, 10mol%), cooled to3° C. and treated with bromofluoromethane (1.10 ml, 19.5 mmol, 0.98equivalents), washing in with prechilled (0° C.) ethyl acetate (EtOAc)(20 ml). The suspension is stirred overnight, allowing to warm to 17° C.The aqueous layer is separated and the organic phase is sequentiallywashed with 1 M HCl (50 ml), 1% w/v NaHCO₃ solution (3×50 ml) and water(2×50 ml). The ethylacetate solution is distilled at atmosphericpressure until the distillate reaches a temperature of approximately 73°C. at which point toluene (150 ml) is added. Distillation is continuedat atmospheric pressure until all remaining EtOAc has been removed(approximate distillate temperature 103° C.). The resultant suspensionis cooled and aged at <10° C. and filtered off. The bed is washed withtoluene (2×30 ml) and the product oven dried under vacuum at 60° C. toconstant weight to yield the title compound (8.77 g, 82%) LCMS retentiontime 3.66min, m/z 539 MH⁺, NMR δ (CDCl₃) includes 7.60 (1H, m),7.18-7.11 (2H, m), 6.52 (1H, dd, J 4.2Hz), 6.46 (1H, s), 6.41 (1H, dd, J10, 2Hz), 5.95 and 5.82 (2H dd, J 51, 9Hz), 5.48 and 5.35 (1H, 2m), 4.48(1H, m), 3.48 (1H, m), 1.55 (3H, s), 1.16 (3H, s), 1.06 (3H, d, J 7Hz).

[0149] Pharmacological Activity

[0150] In Vitro Pharmacological Activity

[0151] Pharmacological activity was assessed in a functional in vitroassay of glucocorticoid agonist activity which is generally predictiveof anti-inflammatory or anti-allergic activity in vivo.

[0152] For the experiments in this section, compound of formula (I) wasused as unsolvated Form 1 (Intermediate 2)

[0153] The functional assay was based on that described by K. P. Ray etal., Biochem J. (1997), 328, 707-715. A549 cells stably transfected witha reporter gene containing the NF-κB responsive elements from the ELAMgene promoter coupled to sPAP (secreted alkaline phosphatase) weretreated with test compounds at appropriate doses for 1 hour at 37° C.The cells were then stimulated with tumour necrosis factor (TNF, 10ng/ml) for 16 hours, at which time the amount of alkaline phosphataseproduced is measured by a standard colourimetric assay. Dose responsecurves were constructed from which EC₅₀ values were estimated.

[0154] In this test the compound of formula (1) showed an EC₅₀value of<1 nM.

[0155] The glucocorticoid receptor (GR) can function in at least twodistinct mechanisms, by upregulating gene expression through the directbinding of GR to specific sequences in gene promotors, and bydownregulating gene expression that is being driven by othertranscription factors (such as NFκB or AP-1) through their directinteraction with GR.

[0156] In a variant of the above method, to monitor these functions, tworeporter plasmids have been generated and introduced separately intoA549 human lung epithelial cells by transfection. The first cell linecontains the firefly luciferase reporter gene under the control of asynthetic promoter that specifically responds to activation of thetranscription factor NFκB when stimulated with TNFα. The second cellline contains the renilla luciferase reporter gene under the control ofa synthetic promotor that comprises 3 copies of the consensusglucocorticoid response element, and which responds to directstimulation by glucocorticoids. Simultaneous measurement oftransactivation and transrepression was conducted by mixing the two celllines in a 1:1 ratio in 96 well plate (40,000 cells per well) andgrowing overnight at 37° C. Test compounds were dissolved in DMSO, andadded to the cells at a final DMSO concentration of 0.7%. Afterincubation for 1 h 0.5 ng/ml TNFα (R&D Systems) was added and after afurther 15 hours at 37° C., the levels of firefly and renilla luciferasewere measured using the Packard Firelite kit following themanufacturers' directions. Dose response curves were constructed fromwhich EC₅₀ values were determined. Transactivation (GR) Transrepression(NFκB) ED₅₀ (nM) ED₅₀ (nM) Compound of Formula 0.06 0.20 (I) Metabolite(X) >250 >1000 Fluticasone propionate 0.07 0.16

[0157] In Vivo Pharmacological Activity

[0158] Pharmacological activity in vivo was assessed in an ovalbuminsensitised Brown Norway rat eosinophilia model. This model is designedto mimic allergen induced lung eosinophilia, a major component of lunginflammation in asthma.

[0159] For the experiments in this section, compound of formula (I) wasused as unsolvated Form 1.

[0160] Compound of formula (I) produced dose dependant inhibition oflung eosinophilia in this model after dosing as an intra-tracheal (IT)suspension in saline 30 min prior to ovalbumin challenge. Significantinhibition is achieved after a single dose of 30 μg of compound offormula (I) and the response was significantly (p=0.016) greater thanthat seen with an equivalent dose of fluticasone propionate in the samestudy (69% inhibition with compound of formula (I) vs 41% inhibitionwith fluticasone propionate).

[0161] In a rat model of thymus involution 3 daily IT doses of 100 μg ofcompound (I) induced significantly smaller reductions in thymus weight(p=0.004) than an equivalent dose of fluticasone propionate in the samestudy (67% reduction of thymus weight with compound (I) vs 78% reductionwith fluticasone propionate).

[0162] Taken together these results indicate a superior therapeuticindex for compound (I) compared to fluticasone propionate.

[0163] In Vitro Metabolism in Rat and Human Hepatocytes

[0164] Incubation of compound (I) with rat or human hepatocytes showsthe compound to be metabolised in an identical manner to fluticasonepropionate with the 17-β carboxylic acid (X) being the only significantmetabolite produced. Investigation of the rate of appearance of thismetabolite on incubation of compound (I) with human hepatocytes (37° C.,10 μM drug concentration, hepatocytes from 3 subjects, 0.2 and 0.7million cells/mL) shows compound (I) to be metabolised ca. 5-fold morerapidly than fluticasone propionate:— 17-β acid metabolite productionSubject Cell density (pmol/h) number (million cells/mL) Compound (I)Fluticasone propionate 1 0.2 48.9 18.8 1 0.7 73.3 35.4 2 0.2 118 9.7 20.7 903 23.7 3 0.2 102 6.6 3 0.7 580 23.9

[0165] Median metabolite production 102-118 pmol/h for compound (I) and18.8-23.0 pmol/h for fluticasone propionate.

[0166] Pharmacokinetics After Intravenous (IV) and Oral Dosing in Rats

[0167] Compound (I) was dosed orally (0.1 mg/kg) and IV (0.1 mg/kg) tomale Wistar Han rats and pharmacokinetic parameters determined. Compound(I) showed negligible oral bioavailability (0.9%) and plasma clearanceof 47.3 mL/min/kg, approaching liver blood flow (plasma clearance offluticasone propionate=45.2 mL/min/kg).

[0168] Pharmacokinetics After Intra-tracheal Dry Powder Dosing in thePig.

[0169] Anaesthetised pigs (2) were dosed intra-tracheally with ahomogenous mixture of compound (I) (1 mg) and fluticasone propionate (1mg) as a dry powder blend in lactose (10% w/w). Serial blood sampleswere taken for up to 8 h following dosing. Plasma levels of compound (I)and fluticasone propionate were determined following extraction andanalysis using LC-MS/MS methodology, the lower limits of quantitation ofthe methods were 10 and 20 pg/mL for compound (I) and fluticasonepropionate respectively. Using these methods compound (I) wasquantifiable up to 2 hours after dosing and fluticasone propionate wasquantifiable up to 8 hours after dosing. Maximum plasma concentrationswere observed for both compounds within 15 min after dosing. Plasmahalf-life data obtained from IV dosing (0.1 mg/kg) was used to calculateAUC (0-inf) values for compound (I). This compensates for the plasmaprofile of Compound (I) only being defined up to 2 hours after an ITdose and removes any bias due to limited data between compound (I) andfluticasone propionate.

[0170] C_(max) and AUC (0-inf) values show markedly reduced systemicexposure to compound (I) compared to fluticasone propionate:— Cmax(pg/mL) AUC (0-inf) (hr. pg/mL) Pig 1 Pig 2 Pig 1 Pig 2 Compound ofFormula (I) 117  81 254 221 Fluticasone propionate 277 218 455 495

[0171] The pharmacokinetic parameters for both compound (I) andfluticasone propionate were the same in the anaesthetised pig followingintravenous administration of a mixture of the two compounds at 0.1mg/kg. The clearance of these two glucocorticoids is similar is thisexperimental pig model.

Intermediate 3:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester, amorphous particles

[0172] Intermediate 2 (30.04 g) was dissolved in methylethylketone (850ml) to give a 3.5% solution. The solution was spray dried using a NiroMobile Minor spray drier (Niro Inc, Columbia, Md., USA). The sprayorifice was a two fluid pneumatic nozzle with 0.04 inch orifice diameter(Spray Systems Co, Wheaton, Ill., USA). The other spray dryingparameters were as follows:

[0173] Temperature: 150° C., outlet temperature 98° C.

[0174] Solution flow rate: 30 ml/min using Isco 260D syringe pump (IscoInc, Lincoln, Nev., USA)

[0175] Atomisation Pressure: 2 Bar

[0176] Particle collection was achieved in the conventional manner usinga Fisher Klosterman XQ120-1.375 high efficiency cyclone(Fisher-Klosterman Inc, Louisville, Ky., USA). A white powder wasrecovered. The spray drying process was successful at producing smooth,spherical particles of amorphous6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester. System yield was 61%

Intermediate 46α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0177] Unsolvated Form 2

EXAMPLES Example 16α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0178] Composition with Toluene

[0179] Intermediate 4 (200 mg) was slurried in toluene (5 mL) for 3hours. The solid was then recovered by filtration to afford the titlecompound as a white solid.

[0180] Stoichiometry of compound of formula (I): guest=1: 3.1 from ¹Hnmr (CDCl₃).

Example 26α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0181] Composition with m-xylene

[0182] Intermediate 4 (200 mg) was slurried in m-xylene (6 mL) for 2hours. The solid was then recovered by filtration to afford the titlecompound as a white solid.

Example 36α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0183] Composition with o-xylene (first alternative method)

[0184] Intermediate 3 (100 mg) was slurried in o-xylene (1 mL) at 21° C.for 16 hours. The solid was recovered by filtration to afford the titlecompound as a white solid. Stoichiometry of compound of formula (I):guest=1: 1.5 from ¹H nmr (CDCl₃).

Example 46α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0185] Composition with o-xylene (second alternative method)

[0186] Intermediate 2 (3.0 g) was dissolved in a mixture of o-xylene (60mL) and acetonitrile (60 mL). The solvent was allowed to evaporate underambient conditions until the crystallisation had proceeded sufficiently.The solid was then recovered by filtration to afford the title compoundas a white solid.

[0187] Stoichiometry of compound of formula (I) : guest=1: 4 from ¹H nmr(CDCl₃).

Example 56α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester

[0188] Composition with Chlorobenzene

[0189] Intermediate 4 (200 mg) was slurried in m-xylene (5 mL) for 3hours. The solid was then recovered by filtration to afford the titlecompound as a white solid.

[0190] Further characterising data on compositions of the invention:

[0191] Detailed XRPD profile peak information for various compositionsof the invention is provided in Tables 2, 3, 4 and 5.

[0192] The XRPD profiles of compositions of the invention are providedin FIGS. 4, 5, 6 and 7.

[0193] We also claim compositions of the invention substantially byreference to their XRPD profiles as shown in the Figures and Tables.

Example A Dry Powder Composition Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester composition with toluene

[0194] A dry powder formulation may be prepared as follows:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β- 0.20 mghydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester, composition with toluene prepared according toExample 1, MMD of 3 μm: milled lactose (wherein not greater than 85% ofparticles 12 mg have a MMD of 60-90 μm, and not less than 15% ofparticles have a MMD of less than 15 μm): A peelable blister stripcontaining 60 blisters each filled with a formulation as just describedmay be prepared.

Example B Dry Powder Composition Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester composition with toluene and a long actingβ₂-adrenoreceptor agonist

[0195] A dry powder formulation may be prepared as follows:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]- 0.20 mg11β-hydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester composition with toluene prepared according toExample 1, MMD of 3 μm: Long-acting β₂-adrenoreceptor agonist(micronised to a 0.20 mg MMD of 3 μm): milled lactose (wherein notgreater than 85% of particles 12 mg have a MMD of 60-90 μm, and not lessthan 15% of particles have a MMD of less than 15 μm): A peelable blisterstrip containing 60 blisters each filled with a formulation as justdescribed may be prepared.

Example C Aerosol Formulation Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester composition with toluene prepared according toExample 1, MMD of 3 μm:

[0196] An aluminium canister may be filled with a fromulation asfollows: 6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β- 250 μghydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester composition with toluene prepared according toExample 1, MMD of 3 μm: 1,1,1,2-tetrafluoroethane: to 50 μl (amounts peractuation) in a total amount suitable for 120 actuations and thecanister may be fitted with a metering valve adapted to dispense 50 μlper actuation.

Example D Aerosol formulation containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester composition with toluene and a long actingβ₂-adrenoreceptor agonist

[0197] An aluminium canister may be filled with a formulation asfollows: 6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β- 250 μghydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester composition with toluene prepared according toExample 1, MMD of 3 μm: Long-acting β₂-adrenoreceptor agonist(micronised to 25 μg a MMD of 3 μm): 1,1,1,2-tetrafluoroethane: to 50 μl(amounts per actuation) in a total amount suitable for 120 actuationsand the canister may be fitted with a metering valve adapted to dispense50 μl per actuation.

Example E Nasal formulation containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester composition with toluene

[0198] A formulation for intranasal delivery may be prepared as follows:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β- 10 mghydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester composition with toluene prepared according toExample 1, MMD of 3 μm: Polysorbate 20 0.8 mg Sorbitan monolaurate 0.09mg Sodium dihydrogen phosphate dihydrate 94 mg Dibasic sodium phosphateanhydrous 17.5 mg Sodium chloride 48 mg Demineralised water to 10 ml Theformulation may be fitted into a spraypump capable of delivering aplurality of metered doses (Valois).

[0199] Throughout the specification and the claims which follow, unlessthe context requires otherwise, the word ‘comprise’, and variations suchas ‘comprises’ and ‘comprising’, will be understood to imply theinclusion of a stated integer or step or group of integers but not tothe exclusion of any other integer or step or group of integers orsteps.

[0200] The patents and patent applications described in this applicationare herein incorporated by reference. TABLE 2 XRPD characteristic anglesand relative intensities for 6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β- carbothioic acidS-fluoromethyl ester, composition with toluene Angle Rel. Intensity2-Theta ° % 4.9 25.6 6.8 12.2 8.0 43.6 9.9 1.7 11.8 15.5 13.0 13.1 14.012.9 15.0 100.0 16.3 53.2 16.7 12.0 17.9 14.2 18.7 83.2 19.4 13.3 20.126.6 21.1 19.7 21.7 17.2 22.8 14.5 23.5 37.2 23.9 34.6 24.5 14.0 25.310.5 25.8 15.5 26.1 20.5 27.0 18.8 28.8 8.2 30.7 39.2 31.1 21.4 32.714.0 34.0 14.0 34.7 18.6

[0201] TABLE 3 XRPD characteristic angles and relative intensities for6α,9α-Difluoro-17α- [(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β- carbothioic acidS-fluoromethyl ester, composition with m-xylene Angle Rel. Intensity2-Theta ° % 4.8 38.7 6.8 12.7 7.9 22.8 9.6 6.2 11.5 24.1 13.0 11.8 13.726.9 14.5 100.0 15.8 17.9 16.1 25.7 16.4 12.4 17.0 7.7 17.6 12.5 18.352.1 19.3 34.9 20.4 6.4 21.2 22.2 22.3 16.3 22.7 18.4 23.3 21.1 24.314.0 25.1 12.8 25.6 14.6 26.3 8.1 26.8 9.5 27.6 6.9 28.0 5.0 28.6 5.829.8 18.8 31.4 8.8 32.5 8.0 33.1 8.7 34.3 11.7

[0202] TABLE 4 XRPD characteristic angles and relative intensities for6α,9α-Difluoro-17α- [(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β- carbothioic acidS-fluoromethyl ester, composition with o-xylene Angle Rel. Intensity2-Theta ° % 5.0 11.4 6.9 13.6 8.1 21.7 9.9 1.6 11.7 18.0 13.1 15.4 13.827.2 14.8 100.0 16.2 39.3 16.6 16.8 17.1 9.4 17.7 12.0 18.5 60.6 19.310.7 19.7 18.5 20.6 5.1 21.5 20.0 22.8 21.3 23.4 22.5 24.4 17.6 24.9 7.425.6 21.5 26.3 12.4 26.8 12.0 28.0 8.8 28.7 7.0 29.0 6.6 30.2 16.6 32.28.1 32.5 9.4 33.4 6.6 34.2 12.3

[0203] TABLE 5 XRPD characteristic angles and relative intensities for6α,9α-Difluoro-17α- [(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β- carbothioic acidS-fluoromethyl ester, composition with chlorobenzene Angle Rel.Intensity 2-Theta ° % 4.3 34.4 6.6 12.4 10.7 10.2 12.7 11.1 13.3 10.513.6 23.4 14.6 22.9 15.3 24.8 16.9 43.2 17.3 44.4 17.6 100.0 19.1 26.719.4 12.4 20.1 32.3 20.9 24.3 22.1 15.2 23.0 47.0 23.6 20.7 24.5 10.525.2 9.6 27.4 12.7 28.5 6.2 29.6 28.1 31.8 17.5 33.3 6.2

1. A crystalline chemical composition comprising a compound of formula(I)

in which the crystal lattice is stabilised by the presence of a guestmolecule, characterised in the crystalline composition is of space groupP2₁2₁2₁ having unit cell dimensions of about 7.7±0.6 Å, 13.7±0.7 Å, and37±3 Å when determined at 120K.
 2. A composition according to claim 1wherein the unit cell dimensions are7.7±0.4 Å, 13.8±0.3 Å, and 36.8±3 Åwhen determined at 120K.
 3. A composition according to claim 1 whereinthe crystal lattice is stabilised by a hydrogen bonding interactionbetween the hydrogen atom of the C11 hydroxy on the compound of formula(I) with the oxygen of the C3 carbonyl on a second molecule of thecompound of formula (I).
 4. A composition according to claim 1 whereinthe guest molecule preferably has a relative molecular weight in therange 16 to
 150. 5. A composition according to claim 1 wherein the guestmolecule is toluene.
 6. A composition according to claim 1 wherein theguest molecule is m-xylene.
 7. A composition according to claim 1wherein the guest molecule is o-xylene.
 8. A composition according toclaim 1 wherein the guest molecule is fluorobenzene.
 9. A compositionaccording to claim 1 wherein the guest molecule is chlorobenzene.
 10. Acomposition according to claim 1 wherein the guest molecule isethylbenzene.
 11. A composition according to claim 1 wherein the ratioof compound of formula (I) to guest molecule is 1:5-0.25.
 12. Apharmaceutical composition comprising a composition according to claim 1together with a physiologically acceptable diluent or carrier.
 13. Amethod for the treatment of a human or animal subject with aninflammatory and/or allergic condition, which method comprisesadministering to said human or animal subject an effective amount of thecomposition according to claim
 1. 14. A pharmaceutical compositioncomprising a composition according to claim 1 in combination withanother therapeutically active agent.
 15. A composition according toclaim 14 wherein the other therapeutically active ingredient is a longacting β₂-adrenoreceptor agonist.
 16. A process for preparing acomposition according to claim 1 which comprises (a) crystallising thecomposition from a solution containing a compound of formula (I) and theguest molecule; or (b) contacting the compound of formula (I) or anothercomposition according claim 1 in solid form with a liquid containing theguest molecule and obtaining the composition therefrom; or (c)contacting a compound of formula (I) or another composition according toclaim 1 in solid form with a vapour containing the guest molecule.